Field of the Invention
The present invention relates generally to the fields of sterile obtention of amniotic fluid and isolation of biomolecules from amniotic fluid. More specifically, the present invention relates to a method to obtain cell free, biologically active and sterile filtered amniotic fluid by high speed centrifugation and membrane filtration.
Description of the Related Art
Within the uterus of a pregnant woman, a growing fetus is surrounded and cushioned by amniotic fluid, a watery liquid within the amnion. Amniotic fluid is one of the main samples used for the medical examination of the pregnant woman and her fetus.
For gathering information about the fetus's health and development, particularly about the possibility of premature birth, amniotic fluid infection, fetal inflammation and infection, fetal damage, fetal maturity, fetal diseases and chromosomal abnormalities, and component analysis of amniotic fluid, amniocentesis (transabdominal amniotic fluid collection) is carried out by inserting a thin, hollow needle through the abdomen into the uterus and taking a small sample of amniotic fluid. Currently, abdominal puncture with a needle for collecting amniotic fluid from the uterus is used for various amniotic fluid tests including the identification of fetal abnormality and amniotic fluid infection. The insertion of the needle, however, entails the risk of complications and medical accidents, causing anxiety and pain to the pregnant woman.
Traditionally, during a Caesarean section, after cutting through the uterus, the amniotic fluid will be suctioned away and discarded to make a bit more room. Amniotic fluid contains cells, electrolytes, growth factors, carbohydrates, lipids, proteins, amino acids, lactate, pyruvate, enzymes and hormones. Amniotic fluid is also a source of stem cells which ideally should be isolated and separately cultivated for cell therapy purposes. While amniotic fluid cells can be obtained from a small amount of fluid during amniocentesis, these amounts are insufficient for a larger scale harvesting of biomolecules or culturing of the stem cells comprising amniotic fluid. Many authors have published that the biological activity of amniotic fluid in many medical applications is the consequence of the presence of the cells, particularly stem cells (Bhattacharya N. Stubblefield P., “Regenerative Medicine Using Pregnancy-Specific Biological Substances” Springer ed. 2011).
Thus, there is a recognized need in the art for an improved means for obtaining sterile amniotic fluid for use in research and the development of therapeutic products. Particularly, the prior art is deficient in methods for obtaining sterile human amniotic fluid with minimal or no risk to a pregnant woman or fetus by collecting the amniotic fluid prior to an elective Caesarean section. Also the prior art is deficient in methods for obtaining sterile human amniotic fluid devoid of cells which may create unwanted reactions due to their allogenic characters on the patients to be treated. The present invention fulfills this longstanding need and desire in the art to improve the safety of amniotic fluid in its medical uses.